论文部分内容阅读
目的:基于肝损伤级联反应关系观察黄药子水煎液(decoction of Dioscorea bulbifera,DDB)连续给药对小鼠肝脏的影响,探讨其致肝损伤的早期作用机制。方法:黄药子水煎液多次灌胃给予小鼠,测定不同给药剂量和不同给药时间下黄药子水煎液对小鼠生化指标、细胞膜指标、氧化与抗氧化指标、线粒体指标、病理组织的影响,并使用Western蛋白印迹法检测小鼠肝组织细胞色素P450(CYP)2E1表达。结果:黄药子水煎液剂量-毒性试验中,较空白对照组2g/kg剂量组小鼠钠钾ATP酶(Na~+-K~+-ATP)、钙镁ATP酶(Ca~(2+)-Mg~(2+)-ATP)、丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽S转移酶(GST)及总巯基(T-SH)均有统计学差异;当剂量增加到20g/kg,小鼠Na~+-K~+-ATP、Ca~(2+)-Mg~(2+)-ATP、MDA、SOD、GST及T-SH均呈显著变化,SDH明显降低。黄药子水煎液时间-毒性试验中,与各自空白对照组比较,小鼠灌胃给予20g/kg剂量的黄药子水煎液,各给药组小鼠丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)均升高,同时黄药子水煎液可显著增强20天给药组及30天给药组CYP2E1表达。10天给药组病理组织学检查可见个别小鼠肝细胞轻微嗜酸变性;20天给药组病理组织学检查可见肝细胞点状或小灶性坏死外,还可见Na~+-K~+-ATP酶活性明显降低;30天给药组小鼠病理组织学检查结果除与20天给药组结果相同外,还可见大量小鼠肝细胞坏死,Na~+-K~+-ATP、Ca~(2+)-Mg~(2+)-ATP酶活性显著降低,同时MDA、SOD、GST、T-SH及SDH均有明显改变。结论:小鼠长期给予DDB可使肝脏发生损伤,损伤程度具有一定的时间和剂量依赖性,其对肝脏的时间-毒性关系为首先降低细胞膜Na~+-K~+-ATP、Ca~(2+)-Mg~(2+)-ATP活性,上调CYP450,进一步破坏氧化与抗氧化系统或线粒体功能,黄药子水煎液致肝损伤早期作用机制可能为损伤肝细胞膜。
OBJECTIVE: To observe the effects of continuous decoction of Dioscorea bulbifera (DDB) on the liver of mice based on the cascade of liver injury and to explore its early mechanism of liver injury. Methods: The mice were given gavage by decoction of Radix Astragali decoction for several times. The effects of Radix Astragali decoction on the biochemical indexes, cell membrane indexes, oxidation and antioxidation indexes, mitochondrial indexes, pathological tissues Western blotting was used to detect the expression of cytochrome P450 (CYP) 2E1 in the liver of mice. Results: Compared with the blank control group, the dose-toxicity test showed that Na + -K + -ATP and Ca 2+ + (P <0.05). There were significant differences in the levels of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione S transferase (GST) and total thiol (T-SH) ; When the dose was increased to 20g / kg, there were significant changes in Na ~ + -K ~ + -ATP, Ca ~ (2 +) - Mg ~ (2 +) - ATP, MDA, SOD, GST and T-SH , SDH decreased significantly. Xanthium decoction time - toxicity test, compared with the respective blank control group, the mice were intragastrically given a dose of 20g / kg xanthones decoction, the mice in each administration group alanine aminotransferase (ALT), aspartate Aminotransferase (AST) were increased, while xanthones decoction could significantly enhance the expression of CYP2E1 in 20-day administration group and 30-day administration group. The histopathological examination of 10 days showed slight slight eosinophilic degeneration of hepatocytes in individual mice. The histopathological examination of 20 days showed that the hepatocytes were punctate or focal necrosis. Na ~ + -K ~ + - ATPase activity was significantly reduced. The histopathological examination results of the mice in the 30-day administration group showed the same results as those of the 20-day administration group, but also a large number of liver cell necrosis, Na ~ + -K ~ + -ATP, Ca ~ (2 +) - Mg ~ (2 +) - ATPase activity was significantly decreased, while MDA, SOD, GST, T-SH and SDH were significantly changed. Conclusion: Long-term administration of DDB in mice can cause damage to the liver in a time-and dose-dependent manner. The time-to-toxicity relationship with the liver is the first to reduce the level of Na ~ + -K ~ + -ATP and Ca ~ +) - Mg ~ (2 +) - ATP, upregulate CYP450, further destroying the oxidation and antioxidant system or mitochondrial function. The early mechanism of hepatic injury induced by Decoction of Radix Astragali may damage the liver cell membrane.